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The document outlines the procedures for mass multiplication of Trichoderma using both solid-state and liquid-state fermentation methods. It details the materials required, step-by-step procedures for inoculation, incubation, and preparation of the final product, including mixing with talcum powder. The solid-state method involves using various substrates, while the liquid-state method utilizes specific broths for cultivation.

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4 views1 page

PPATH

The document outlines the procedures for mass multiplication of Trichoderma using both solid-state and liquid-state fermentation methods. It details the materials required, step-by-step procedures for inoculation, incubation, and preparation of the final product, including mixing with talcum powder. The solid-state method involves using various substrates, while the liquid-state method utilizes specific broths for cultivation.

Uploaded by

poojaintudyog
Copyright
© All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PDF, TXT or read online on Scribd

II.

Mass multiplication of Trichoderma


A. Trichoderma under solid-state fermentation (talc based formulation)
Materials required : sorghum, wheat straw, wheat bran, sawdust, coffee husk, sand corn meal
medium, rice bran, vegetable waste, 250 ml Erlenmayer flask, Inoculation needle, Trichoderma culture,
Talcum powder, CMC, sieves, grinder, polypropylene bags.
Procedure:
1. Wash and soak the grains/substrate in water for 12 hrs and then filled in 250 ml Erlenmayer flasks (@
100g/ flask) or in half kg polypropylene bags (150 g / bag)
2. Plug the flasks and bags with non-absorbent cotton plug (with the help of neck)
3. Autoclave at 20 lbs. psi. for 20 min.
4. After cooling to room temperature, inoculate the flasks or bags with 4-6 disc of actively growing
culture of Trichoderma spp.
5. Incubate the bags at 28 0C for 10-12 days and shake at regular intervals.
6. The colonized grains turn green with sporulation
7. After 12 days, the colonized grains are emptied into plastic trays and dried under shade for 4-5 days.
8. The dried colonized grains are then ground with the help of Willey Mill or a normal grinder to get a
powder
9. This powder is then sieved through a normal coarse (50 mesh size) and fine (80 mesh size) sieves,
simultaneously to obtain a very fine pure powder
[Link] pure powder is then mixed with talcum powder in a ratio of 10 g pure powder + 1 kg talc powder
(commercial grade) to get the commercial formulation
11.10 % Carboxy Methyl Cellulose (CMC) is also added as a sticker

[Link] under liquid-state fermentation


Material required: Czapek dox broth, V8 broth, Richards broth/ PD broth/ molasses brewer’s yeast
medium/ molasses soy medium/jaggery medium
Procedure:
1. Prepare molasses brewer’s yeast medium (molasses – 30 g, yeast powder – 5 g, distilled water-1 lit.)
by mixing the ingredients of respective medium in conical flasks/ glass bottles and autoclave them.
2. After cooling those at room temperature inoculate with mycelia and spore discs from mother culture.
3. Inoculate the flasks at room temperature either as stationary culture or on a rotary shaker at 150-180
rpm for 3-5 days.
4. Keep at a stationary stage for 1-2 days and obtain the mycelia mat and spores to make the
formulation after drying or directly mixing it with talc powder.

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